Pandanus amaryllifolius is a member of the family Pandanaceae comprising a group of plants generally known as ‘screw pines’. Earlier studies by Buttery et al (Chem. Ind. London, Vol. 23, page 478 of year 1982) indicated that 2-acetyl-1-pyrroline is the major flavour constituent of Pandanus leaves, while other constituents like volatile oils, alcohols, aromatic aldehydes, ketones and esters were also traceable. The major compound, 2-acetyl-1-pyrroline, has commercial significance as a major flavour constituent that impart the characteristic aroma of scented rice varieties such as Basmati rice of India, ‘Khao Dawk Mali-105’ of Thailand and Kaorimai of Japan. The essential oil of Pandanus is known to impart 10-times more flavour than the highly scented rice and 100-times more than that present in normal rice (Chem. Ind. London, Vol. 23, page 478 of year 1982). Pandanus extract has been reported to possess anti-oxidant properties. Pandanus leaf is cited as an ingredient in many Indian, Thai and Indonesian recipes. Pandanus amaryllifolius is not known to grow in wild state whereas the plants are grown as ornamentals in pots or grown in kitchen gardens in parts of South East Asia, Indonesia and New Guinea. The plant produces only 6-8 suckers per year and is known to produce male flowers only in New Guinea. Female flowers are not known in the variety that synthesizes aroma in the leaves. Thus the lack of its wild population or the large-scale cultivation on one hand and a high potential for its economic importance on the other hand indicate a huge future demand for this plant material. Thus there could be possibilities of its organized cultivation and hence would be a demand for the planting material. To meet such demands, a method for rapid production of plantlets by tissue culture means has tremendous scope. Since the flavor compound in this plant is known to vary in each variety, the plant material with highest perceivable flavor level was selected and a method for rapid multiplication of plantlets was invented.
Plant Tissue Culture (PTC) is a major area of plant biotechnology, which has direct impact on present day agriculture/horticulture. The need for enormous supply of desired type of planting/sowing material may be obtained by applying tissue culture method as an alternative to conventional method. At present, several laboratories in the world are producing over 650 million plants annually, by applying tissue culture method.
There are various advantages of tissue culture (TC) plants over those produced by conventional methods of plant propagation. As TC plants are produced in completely controlled environment, the chances of carrying systemic diseases are rare. The plants are induced to multiply at a tremendous rate by developing specific medium formulation for each plant material. Maximum number of plants can be produced using minimum space, time and nutrients. Similarly, storage and transportation can be handled with lesser cost than for conventional plants. It facilitates the availability of planting material throughout the year.
The present invention relates to a medium formulation for the clonal propagation of scented Pandanus amaryllifolius Roxb. wherein the development of specific nutrient medium with growth regulators help establishment of shoot cultures and rapid further aseptic multiplication of Pandanus shoots which may subsequently be hardened to obtain high quality Pandanus planting material.
There are no reports on the tissue culture studies or clonal propagation of Pandanus amaryllifolius. However there are certain studies on the chemical composition of the plant at different stages of its life cycle.
Laksanamai V. and Ilangantilake S. (Cereal Chemistry Vol. 70 (4) 1983) found 2-acetyl-1-pyrroline in Pandanus leaves at levels 1 ppm (on fresh weight basis) and described this aroma as that present in scented rice varieties.
Another flavour, ethyl formamide present in rice as a flavouring compound has been identified in Pandanus leaves (Naturwissenchaften, Vol. 71: 215, 1984). Yet another study found 3-methyl-2-(5H)-furanone as the main volatile compound in Pandanus leaves, besides 3-hexanol, 4-methylpentanol, 3-hexanone and 2-hexanone (Flavour Chemistry of ethnic foods—Proceedings of a Meeting held during the 5th Chemical Congress of North America, Cancun, Nov. 11-15, 1997 (1999)).
The drawback associated with the methods followed hitherto for the supply of planting material is that of the conventional type of propagation involves injury to the mother plant for collecting the lateral shoot suckers which is often associated with the sacrificing of the selected high yielding mother plant. Conventional method of propagation transmits systemic infection present in the mother plant acquired from one generation to the next leading to diseased planting material. The rate of multiplication is slow.